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anti-phospho-pyk2 py402 antibody sc-293142 (Santa Cruz Biotechnology)

Name: anti-phospho-pyk2 py402 antibody sc-293142
Brand: Santa Cruz Biotechnology
Rating: 90 (1 reviews)

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Santa Cruz Biotechnology anti-phospho-pyk2 py402 antibody sc-293142
Anti Phospho Pyk2 Py402 Antibody Sc 293142, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-phospho-pyk2 py402 antibody sc-293142/product/Santa Cruz Biotechnology
Average 90 stars, based on 1 article reviews

anti-phospho-pyk2 py402 antibody sc-293142 - by Bioz Stars, 2026-02

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( A ) Representative bright-field pictures of wildtype (WT) and S100a9 -/- neutrophils spreading over E-selectin, ICAM-1, and CXCL1 coated glass capillaries (scale bars = 10 μm). Analysis of cell shape parameters ( B ) area, perimeter, ( C ) circularity (4π * [area/perimeter]) and solidity (area/convex area) over time (mean + SEM, n = 103 [WT] and 96 [ S100a9 -/- ] neutrophils of 4 mice per group, unpaired Student’s t-test). ( D ) Rose plot diagrams representative of migratory crawling trajectories of WT and S100a9 -/- neutrophils in flow chambers coated with E-selectin, ICAM-1, and CXCL1 under flow (2 dyne cm –2 ). Analysis of ( E ) crawling distance, ( F ) directionality of migration, and ( G ) crawling velocity of WT and S100a9 -/- neutrophils (mean + SEM, n = 5 mice per group, 113 [WT] and 109 [ S100a9 -/- ] cells, paired Student’s t-test). Western blot analysis of ICAM-1-induced ( H ) Pyk2 and ( I ) paxillin phosphorylation of WT and S100a9 -/- neutrophils upon CXCL1 stimulation (10 nM) (mean + SEM, representative western blot of n ≥ 4 mice per group, two-way analysis of variance (ANOVA), Sidak’s multiple comparison). ns, not significant; *p≤0.05, **p≤0.01, ***p≤0.001. Figure 3—source data 1. Pyk2 and paxillin western blot data for . Original membranes corresponding to . paxillin, p-paxillin, Pyk2, and p-Pyk2 membranes are depicted and representative blots were then cropped and edited. Rectangle boxes indicate the representative bands used in the figure. Lowest membranes were cut before the staining to save staining solution and fit more membranes at the same time. GAPDH was always employed as an internal control. Chamaleon Duo Pre-stained Protein Ladder was used as molecular weight marker. Figure 3—source data 2. Pyk2 and paxillin western blot data for . Original membranes corresponding to . Paxillin, p-paxillin, Pyk2, and p-Pyk2 original membranes.

Journal: eLife

Article Title: Cytosolic S100A8/A9 promotes Ca 2+ supply at LFA-1 adhesion clusters during neutrophil recruitment

doi: 10.7554/eLife.96810

Figure Lengend Snippet: ( A ) Representative bright-field pictures of wildtype (WT) and S100a9 -/- neutrophils spreading over E-selectin, ICAM-1, and CXCL1 coated glass capillaries (scale bars = 10 μm). Analysis of cell shape parameters ( B ) area, perimeter, ( C ) circularity (4π * [area/perimeter]) and solidity (area/convex area) over time (mean + SEM, n = 103 [WT] and 96 [ S100a9 -/- ] neutrophils of 4 mice per group, unpaired Student’s t-test). ( D ) Rose plot diagrams representative of migratory crawling trajectories of WT and S100a9 -/- neutrophils in flow chambers coated with E-selectin, ICAM-1, and CXCL1 under flow (2 dyne cm –2 ). Analysis of ( E ) crawling distance, ( F ) directionality of migration, and ( G ) crawling velocity of WT and S100a9 -/- neutrophils (mean + SEM, n = 5 mice per group, 113 [WT] and 109 [ S100a9 -/- ] cells, paired Student’s t-test). Western blot analysis of ICAM-1-induced ( H ) Pyk2 and ( I ) paxillin phosphorylation of WT and S100a9 -/- neutrophils upon CXCL1 stimulation (10 nM) (mean + SEM, representative western blot of n ≥ 4 mice per group, two-way analysis of variance (ANOVA), Sidak’s multiple comparison). ns, not significant; *p≤0.05, **p≤0.01, ***p≤0.001. Figure 3—source data 1. Pyk2 and paxillin western blot data for . Original membranes corresponding to . paxillin, p-paxillin, Pyk2, and p-Pyk2 membranes are depicted and representative blots were then cropped and edited. Rectangle boxes indicate the representative bands used in the figure. Lowest membranes were cut before the staining to save staining solution and fit more membranes at the same time. GAPDH was always employed as an internal control. Chamaleon Duo Pre-stained Protein Ladder was used as molecular weight marker. Figure 3—source data 2. Pyk2 and paxillin western blot data for . Original membranes corresponding to . Paxillin, p-paxillin, Pyk2, and p-Pyk2 original membranes.

Article Snippet: After subsequent blocking (LI-COR blocking solution), membranes were incubated with the following antibodies for later detection and analysis using the Odyssey CLx Imaging System and Image Studio software: rabbit α-mouse phospho-paxillin (Tyr118) or rabbit α-mouse paxillin and rabbit α-mouse phospho-Pyk2 (Tyr402) or rabbit α-mouse Pyk2 (all Cell Signaling).

Techniques: Migration, Western Blot, Phospho-proteomics, Comparison, Staining, Control, Molecular Weight, Marker

Ca V 1.2/1.3 regulates IDO1 expression via Pyk2 and calmodulin. A,B) MDA‐MB‐231 cells were pre‐treated with lacidipine (10 × 10 −6 , 20 × 10 −6 , and 40 × 10 −6 m ) and tyrphostin A9 (5 × 10 −6 m ) or W‐7 (35 × 10 −6 m ) for 2 h, then stimulated with IFN γ (100 ng mL −1 ) for 24 h. The expression of Pyk2 phosphorylated at A) Tyr402, Pyk2 and B) calmodulin (B) proteins was analyzed by western blotting. GAPDH was used as the loading control. MDA‐MB‐231 cells were pre‐treated with lacidipine (20 × 10 −6 m ) and C) tyrphostin A9 (5 × 10 −6 m ) or D) W‐7 for 2 h, then stimulated with IFN γ (100 ng mL −1 ) for 24 h. The expression of IDO1 proteins was analyzed by western blotting. GAPDH was used as the loading control. MDA‐MB‐231 cells were pre‐treated with E) lacidipine (20 × 10 −6 m ) and tyrphostin A9 (5 × 10 −6 m ) or F) W‐7 for 2 h, then stimulated with IFN γ (100 ng mL −1 ) for 24 h. The expression of phosphorylated JAK at Tyr1022, phosphorylated STAT1 at Tyr701, phosphorylated IKK α/β, phosphorylated IκBα, and IκBα proteins was analyzed by western blotting. GAPDH was used as the loading control. G) MDA‐MB‐231 cells were treated with lacidipine (20 × 10 −6 m ) for 3 h and then co‐immunoprecipitated with Pyk2 antibodies or control immunoglobulin G (IgG) and analyzed for antibody‐specific JAK1, calmodulin, and Pyk2. The MDA‐MB‐231 cells were transfected with the indicated H) Ca V 1.2 and I) Ca V 1.3 siRNA. After transfection for 48 h, cells were treated with IFN γ (100 ng mL −1 ) for 24 h and cell lysates were collected for western blot analysis using the indicated antibodies to assess the effect of Ca V 1.2 and Ca V 1.3. GAPDH was used as the loading control. All experiments were conducted with three independent replicates.

Journal: Advanced Science

Article Title: Calcium Channel Blocker Lacidipine Promotes Antitumor Immunity by Reprogramming Tryptophan Metabolism

doi: 10.1002/advs.202409310

Figure Lengend Snippet: Ca V 1.2/1.3 regulates IDO1 expression via Pyk2 and calmodulin. A,B) MDA‐MB‐231 cells were pre‐treated with lacidipine (10 × 10 −6 , 20 × 10 −6 , and 40 × 10 −6 m ) and tyrphostin A9 (5 × 10 −6 m ) or W‐7 (35 × 10 −6 m ) for 2 h, then stimulated with IFN γ (100 ng mL −1 ) for 24 h. The expression of Pyk2 phosphorylated at A) Tyr402, Pyk2 and B) calmodulin (B) proteins was analyzed by western blotting. GAPDH was used as the loading control. MDA‐MB‐231 cells were pre‐treated with lacidipine (20 × 10 −6 m ) and C) tyrphostin A9 (5 × 10 −6 m ) or D) W‐7 for 2 h, then stimulated with IFN γ (100 ng mL −1 ) for 24 h. The expression of IDO1 proteins was analyzed by western blotting. GAPDH was used as the loading control. MDA‐MB‐231 cells were pre‐treated with E) lacidipine (20 × 10 −6 m ) and tyrphostin A9 (5 × 10 −6 m ) or F) W‐7 for 2 h, then stimulated with IFN γ (100 ng mL −1 ) for 24 h. The expression of phosphorylated JAK at Tyr1022, phosphorylated STAT1 at Tyr701, phosphorylated IKK α/β, phosphorylated IκBα, and IκBα proteins was analyzed by western blotting. GAPDH was used as the loading control. G) MDA‐MB‐231 cells were treated with lacidipine (20 × 10 −6 m ) for 3 h and then co‐immunoprecipitated with Pyk2 antibodies or control immunoglobulin G (IgG) and analyzed for antibody‐specific JAK1, calmodulin, and Pyk2. The MDA‐MB‐231 cells were transfected with the indicated H) Ca V 1.2 and I) Ca V 1.3 siRNA. After transfection for 48 h, cells were treated with IFN γ (100 ng mL −1 ) for 24 h and cell lysates were collected for western blot analysis using the indicated antibodies to assess the effect of Ca V 1.2 and Ca V 1.3. GAPDH was used as the loading control. All experiments were conducted with three independent replicates.

Article Snippet: The embedded tissues were cut into 4 × 10 −6 m slices and incubated with the antibodies against IDO1 (#13268‐1‐AP, ProteinTech), Pyk2 (Phospho‐Tyr402) (#11216, Signalway), CD8 (#47992, Signalway), and Foxp3 (#AF6544, Affinity) overnight at 4 °C.

Techniques: Expressing, Western Blot, Control, Immunoprecipitation, Transfection

The expression levels of IDO1 and Pyk2 phosphorylation correlate with human breast cancer development. A) Statistical analysis of IDO1 expression in the breast tumor tissues ( n = 136). B,C) Expression of IDO1 in different subtypes of breast cancer ( n = 136). D) Statistical analysis of Pyk2 phosphorylation expression in the breast tumor tissues ( n = 136). E,F) Expression levels of Pyk2 phosphorylation in different subtypes of breast cancer ( n = 136). G) The correlation of IDO1 and Pyk2 phosphorylation, R = 0.3, p = 0.00042. H) Expression levels of IDO1 in the breast tumor tissues, adjacent normal breast tissues, and normal breast tissue based on TCGA data ( n = 1034). I) Expression levels of IDO1 in different subtypes of breast cancer based on TCGA data, p < 0.001 ( n = 1034).

Journal: Advanced Science

Article Title: Calcium Channel Blocker Lacidipine Promotes Antitumor Immunity by Reprogramming Tryptophan Metabolism

doi: 10.1002/advs.202409310

Figure Lengend Snippet: The expression levels of IDO1 and Pyk2 phosphorylation correlate with human breast cancer development. A) Statistical analysis of IDO1 expression in the breast tumor tissues ( n = 136). B,C) Expression of IDO1 in different subtypes of breast cancer ( n = 136). D) Statistical analysis of Pyk2 phosphorylation expression in the breast tumor tissues ( n = 136). E,F) Expression levels of Pyk2 phosphorylation in different subtypes of breast cancer ( n = 136). G) The correlation of IDO1 and Pyk2 phosphorylation, R = 0.3, p = 0.00042. H) Expression levels of IDO1 in the breast tumor tissues, adjacent normal breast tissues, and normal breast tissue based on TCGA data ( n = 1034). I) Expression levels of IDO1 in different subtypes of breast cancer based on TCGA data, p < 0.001 ( n = 1034).

Techniques: Expressing

Fig. 1 Inhibition of Pyk2 signaling induces multinucleation formation in microglia. A Morphological changes between mononuclear and multinucleated microglia. Immunocytochemistry of MG6 cells stained with anti-Iba1 (green) and anti-β-tubulin (magenta). Nuclear DNA was labeled with DAPI (white). Scale bar is 20 μm. B Cells were treated with Pyk2-Inh for 24 h. Phase contrast images (upper panel) and DAPI stained images (lower panel) were shown. Circles are indicated as multinucleated cells. C The quantification of the number of multinucleated cells/well (control: n = 4, Pyk2-Inh-500 nM: n = 4, Pyk2-Inh-1000 nM). D Cell proliferation was assessed by CCK-8. MG6 cells were treated with 100 μg/mL CSF1 and indicated concentration of Pyk2-Inh (n = 6 per group, One-way ANOVA, p < 0.05). E Representatives immunoblot images. The ratio of p-Pyk2/ Pyk2, pFAK/FAK, p-ERK/ERK, p-Src/Src and Mannose receptor/Tubulin were described under the blot. Pyk2-Inh was administered as a pre-treatment two hours before stimulation with CSF1 (100 μg/mL). F Image quantification and analysis were performed. The same experiments were repeated three times. All full-length uncropped original western blots are included in a Sup. Fig. 4. Values are mean ± SD. #: negative control (without CSF1) vs control (with CSF1), ##: p < 0.01; *: control (with CSF1) vs Pyk2-Inh (with CSF1), *: p < 0.05; **: p < 0.01; ***: p-value < 0.001; ****: p-value < 0.0001

Journal: Journal of neuroinflammation

Article Title: Enhanced phagocytosis associated with multinucleated microglia via Pyk2 inhibition in an acute β-amyloid infusion model.

doi: 10.1186/s12974-024-03192-7

Figure Lengend Snippet: Fig. 1 Inhibition of Pyk2 signaling induces multinucleation formation in microglia. A Morphological changes between mononuclear and multinucleated microglia. Immunocytochemistry of MG6 cells stained with anti-Iba1 (green) and anti-β-tubulin (magenta). Nuclear DNA was labeled with DAPI (white). Scale bar is 20 μm. B Cells were treated with Pyk2-Inh for 24 h. Phase contrast images (upper panel) and DAPI stained images (lower panel) were shown. Circles are indicated as multinucleated cells. C The quantification of the number of multinucleated cells/well (control: n = 4, Pyk2-Inh-500 nM: n = 4, Pyk2-Inh-1000 nM). D Cell proliferation was assessed by CCK-8. MG6 cells were treated with 100 μg/mL CSF1 and indicated concentration of Pyk2-Inh (n = 6 per group, One-way ANOVA, p < 0.05). E Representatives immunoblot images. The ratio of p-Pyk2/ Pyk2, pFAK/FAK, p-ERK/ERK, p-Src/Src and Mannose receptor/Tubulin were described under the blot. Pyk2-Inh was administered as a pre-treatment two hours before stimulation with CSF1 (100 μg/mL). F Image quantification and analysis were performed. The same experiments were repeated three times. All full-length uncropped original western blots are included in a Sup. Fig. 4. Values are mean ± SD. #: negative control (without CSF1) vs control (with CSF1), ##: p < 0.01; *: control (with CSF1) vs Pyk2-Inh (with CSF1), *: p < 0.05; **: p < 0.01; ***: p-value < 0.001; ****: p-value < 0.0001

Article Snippet: The primary antibodies detected immuno-blotting were listed as follows: p-Pyk2 (Anti-rabbit, Cell Signaling Technology #3291, 1:1000), Pyk2 (Anti-rabbit, Cell Signaling Technology #3292, 1:1000), pFAK (Anti-rabbit, Cell Signaling Technology #3283, 1:1000), FAK (Anti-Rabbit, Cell Signaling Technology #3285, 1:1000), p-ERK (Anti-Rabbit, Cell Signaling Technology #4370, 1:1000), ERK (Anti-Rabbit, Cell Signaling Technology #9102, 1:1000), p-Src (Anti-rabbit, Cell Signaling Technology #6943, 1:1000), Src (Antirabbit, Cell Signaling Technology #2108, 1:1000), Mannose receptor (Anti-mouse, Abcam ab64693, 1:1000), Lamp-1 (Anti-rabbit, Abcam ab24170, 1:1000), p-NF-κB (Anti-rabbit, Cell Signaling Technology #3033, 1:1000), NF-κB (Anti-rabbit, Cell Signaling Technology #8242, 1:1000), β-actin (Anti-mouse, Wako, 010-27841, 1:1000) and Tubulin (Anti-rat, Abcam ab6160, 1:1000).

Techniques: Inhibition, Immunocytochemistry, Staining, Labeling, Control, CCK-8 Assay, Concentration Assay, Western Blot, Negative Control

Fig. 2 Multinucleated cells are generated from abscission failure in cytokinesis, not cell fusion. A Dynamics of cell division by time-lapse video microscopy. The phase contrast images of the microglia’s mitosis at different times. Nuclei were observed with Hoechst 33,342 staining. Arrowheads indicate representative images from anaphase to cytokinesis (control: n = 4, Pyk2-Inh: n = 4). B Cell division was quantified, categorizing it as either appropriate or inappropriate, in both control (n = 4) and Pyk2-Inh treated groups (n = 4). C Fluorescence images of microglia cells showing the stages of cell division. Immunocytochemistry of MG6 cells stained with anti-Pericentrin (red), anti-β-tubulin (green) and phalloidin (white). Nuclear DNA was labeled with DAPI (blue). Independent experiments were performed at least three times

Journal: Journal of neuroinflammation

Article Title: Enhanced phagocytosis associated with multinucleated microglia via Pyk2 inhibition in an acute β-amyloid infusion model.

doi: 10.1186/s12974-024-03192-7

Figure Lengend Snippet: Fig. 2 Multinucleated cells are generated from abscission failure in cytokinesis, not cell fusion. A Dynamics of cell division by time-lapse video microscopy. The phase contrast images of the microglia’s mitosis at different times. Nuclei were observed with Hoechst 33,342 staining. Arrowheads indicate representative images from anaphase to cytokinesis (control: n = 4, Pyk2-Inh: n = 4). B Cell division was quantified, categorizing it as either appropriate or inappropriate, in both control (n = 4) and Pyk2-Inh treated groups (n = 4). C Fluorescence images of microglia cells showing the stages of cell division. Immunocytochemistry of MG6 cells stained with anti-Pericentrin (red), anti-β-tubulin (green) and phalloidin (white). Nuclear DNA was labeled with DAPI (blue). Independent experiments were performed at least three times

Techniques: Generated, Microscopy, Staining, Control, Fluorescence, Immunocytochemistry, Labeling

Fig. 5 Altered morphological features and phagocytic function in microglia of Iba-1 EGFP Tg mice brain after Pyk2-Inh infusion. A The experimental schedule for brain infusion assay in Iba1-EGFP Tg 7-week-old mice (n = 3/group). B Confocal images of GFP positive microglia (green) and β-amyloid (red). Obtained brain slices (thickness ≤ 1.0 mm) were treated with RapiClear 1.52 for tissue clearing and immunohistochemical assay was conducted. C, D Morphological changes were analyzed by IMARIS software. Cell surface, dendrite volume, and dendrite length were quantified. E Spot analysis by IMARIS software. F, G Quantification of the number of β-amyloid spots per cell and number of microglia deposits. H Representative confocal image of multinucleated microglia (indicated red arrow). Brain slices were stained with anti-NeuN (white) and nuclear DNA was stained with DAPI (white). Values are mean ± SD. *: p < 0.05; **: p < 0.01; ***: p-value < 0.001; ****: p-value < 0.0001

Journal: Journal of neuroinflammation

Article Title: Enhanced phagocytosis associated with multinucleated microglia via Pyk2 inhibition in an acute β-amyloid infusion model.

doi: 10.1186/s12974-024-03192-7

Figure Lengend Snippet: Fig. 5 Altered morphological features and phagocytic function in microglia of Iba-1 EGFP Tg mice brain after Pyk2-Inh infusion. A The experimental schedule for brain infusion assay in Iba1-EGFP Tg 7-week-old mice (n = 3/group). B Confocal images of GFP positive microglia (green) and β-amyloid (red). Obtained brain slices (thickness ≤ 1.0 mm) were treated with RapiClear 1.52 for tissue clearing and immunohistochemical assay was conducted. C, D Morphological changes were analyzed by IMARIS software. Cell surface, dendrite volume, and dendrite length were quantified. E Spot analysis by IMARIS software. F, G Quantification of the number of β-amyloid spots per cell and number of microglia deposits. H Representative confocal image of multinucleated microglia (indicated red arrow). Brain slices were stained with anti-NeuN (white) and nuclear DNA was stained with DAPI (white). Values are mean ± SD. *: p < 0.05; **: p < 0.01; ***: p-value < 0.001; ****: p-value < 0.0001

Techniques: Immunohistochemical staining, Software, Staining

Fig. 6 Pyk2 inhibition reduces LPS-induced inflammation in human microglia. A The transcriptional expression of IL1β, IL6 and TNF in control and Pyk2-Inh treated group in human microglia cells, HMC3. B The secreted protein level of IL-6 was measured by ELISA. C The time-course Immunoblotting images of phosphor-NF-kB and NF-kB in HMC3 cells after LPS stimulation and with or without Pyk2-Inh. β-actin was used for loading control. The same experiments were repeated three times. D Quantification of immunoblotting data was measured by ImageJ. All full-length uncropped original western blots are included in a Sup. Fig. 6. Values are mean ± SD. #: significance between negative control and LPS control, #: p < 0.05; ##: p < 0.01; *: significance between LPS control and Pyk2-Inh treated groups, *: p < 0.05; **: p < 0.01; ***: p < 0.001; ns: not significant

Journal: Journal of neuroinflammation

Article Title: Enhanced phagocytosis associated with multinucleated microglia via Pyk2 inhibition in an acute β-amyloid infusion model.

doi: 10.1186/s12974-024-03192-7

Figure Lengend Snippet: Fig. 6 Pyk2 inhibition reduces LPS-induced inflammation in human microglia. A The transcriptional expression of IL1β, IL6 and TNF in control and Pyk2-Inh treated group in human microglia cells, HMC3. B The secreted protein level of IL-6 was measured by ELISA. C The time-course Immunoblotting images of phosphor-NF-kB and NF-kB in HMC3 cells after LPS stimulation and with or without Pyk2-Inh. β-actin was used for loading control. The same experiments were repeated three times. D Quantification of immunoblotting data was measured by ImageJ. All full-length uncropped original western blots are included in a Sup. Fig. 6. Values are mean ± SD. #: significance between negative control and LPS control, #: p < 0.05; ##: p < 0.01; *: significance between LPS control and Pyk2-Inh treated groups, *: p < 0.05; **: p < 0.01; ***: p < 0.001; ns: not significant

Techniques: Inhibition, Expressing, Control, Enzyme-linked Immunosorbent Assay, Western Blot, Negative Control

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